We conducted an expirement to detect disulphide groups in protein. One tube(lysozyme) contained unkowns, had approx 9.34 micromoles of disulphide groups.

The question asks me to work backwards through all the dilution factors taken to give number of micromoles of disulphide groups per ml of original solution.

These are the steps taken to prepare the lysozyme tube:
- A tube of lysozyme solution of 1:32 dilution of lysozyme solution( 0.5mg/ml)
0.0001M of FMA in 0.01M NAOH (provided) should be diluted 1:10 with 1M NaOH.
-Add 8ml of 1M NaOH

Please if you any further questions, let me know
thanks

To work backwards and determine the concentration of disulphide groups in the original solution, we need to consider the dilution factors.

1. Initial dilution: The lysozyme tube is prepared by diluting the original solution by a factor of 1:32. This means that 1 part of the original solution is mixed with 32 parts of the diluent (in this case, probably a buffer or solvent).

2. FMA dilution: The FMA solution is provided at a concentration of 0.0001M in 0.01M NaOH. It needs to be further diluted with 1M NaOH. Let's assume you used a certain volume of the FMA solution and added 8ml of 1M NaOH to it.

To calculate the dilution factor for FMA, we need to know the volume of the FMA solution used before adding the 8ml of 1M NaOH. Let's call this volume "V" (in ml).

The final volume after adding 8ml of 1M NaOH will be V + 8ml. So the dilution factor will be (V + 8) / V.

3. Determination of disulphide group concentration: Based on the information provided, there are roughly 9.34 micromoles of disulphide groups in the lysozyme tube. This concentration is per milliliter of the lysozyme tube.

Therefore, to calculate the concentration of disulphide groups in the original solution (per ml), you need to multiply the concentration in the lysozyme tube by the inverse of the dilution factor at each step.

For example, if you used 1 ml of the FMA solution before adding 8ml of 1M NaOH, the dilution factor for FMA would be (1 + 8) / 1 = 9.

To calculate the concentration of disulphide groups per ml in the original solution, you would divide 9.34 micromoles/ml (from the lysozyme tube) by the dilution factor at each step.

Remember to consider the dilution factors for both the lysozyme dilution and the FMA dilution before reaching the final answer.

Please provide the actual volume of the FMA solution used, and I can help you work through the calculation.